Transcription

1 A RT I C L E S Obesity alters the lung myeloid cell landscape to enhance breast cancer metastasis through IL and GM-CSF Daniela F. Quail,7, Oakley C. Olson,7, Priya Bhardwaj, Logan A. Walsh, Leila Akkari,,, Marsha L. Quick, I-Chun Chen, Nils Wendel, Nir Ben-Chetrit,, Jeanne Walker, Peter R. Holt, Andrew J. Dannenberg and Johanna A. Joyce,,,8 Obesity is associated with chronic, low-grade inflammation, which can disrupt homeostasis within tissue microenvironments. Given the correlation between obesity and relative risk of death from cancer, we investigated whether obesity-associated inflammation promotes metastatic progression. We demonstrate that obesity causes lung neutrophilia in otherwise normal mice, which is further exacerbated by the presence of a primary tumour. The increase in lung neutrophils translates to increased breast cancer metastasis to this site, in a GM-CSF- and IL-dependent manner. Importantly, weight loss is sufficient to reverse this effect, and reduce serum levels of GM-CSF and IL in both mouse models and humans. Our data indicate that special consideration of the obese patient population is critical for effective management of cancer progression. Tumours develop in complex microenvironments containing diverse cell types and inflammatory mediators,. Beyond the local tumour microenvironment, an inflammatory systemic environment can also affect disease outcome, by perturbing homeostasis within multiple tissues throughout the body. This becomes particularly important during metastasis, where systemic alterations can modify the tissue landscape of distant organs and support tumour cell colonization by establishing a pre-metastatic niche. Indeed, chronic inflammation can increase cancer risk and/or progression. Investigation into how the systemic environment affects tumour biology is therefore critical for an integrated understanding of cancer. A prevalent and clinically relevant example of systemic inflammation is obesity, which affects >% of adults in the US, and is linked to multiple pathologies, including cancer. As a growing epidemic, obesity now rivals smoking as the leading preventable cause of cancer 7. Obesity-associated inflammation is driven in part by adipocyte myeloid cell interactions within various fat depots, resulting in altered immune cell composition in different tissues 8. Thus, an emerging hypothesis is that obesity-associated inflammation promotes cancer progression, aligning with well-established associations between other chronic inflammatory conditions and tumorigenesis. Clinical analyses have shown that obesity is associated with higher incidence of breast cancer metastasis, particularly to liver and lung. However, insight into the mechanisms underlying obesity-induced metastasis, compared with the effects on primary tumour growth, has been limited. Of particular interest is lung metastasis, given the high frequency of breast cancer dissemination to this site in patients 7, and the strong clinical association between obesity and multiple lung-inflammatory conditions 8,9, despite the lack of adipose tissue within the lung. Therefore, we sought to analyse the effects of obesity-associated inflammation on breast-to-lung metastasis using animal models, focusing on myeloid cell populations given their role during adipose inflammation and pulmonary metastasis in nonobese settings 9. RESULTS Obesity is associated with lung neutrophilia driven by high adiposity To investigate the effects of obesity on lung inflammation, we used a diet-induced obesity (DIO) model; wild-type () C7 black (BL) female mice were enrolled on either a low-fat (; % kcal) or highfat (; % kcal) diet for weeks followed by immunoprofiling. We found lungs exhibited elevated proportions of CD + leukocytes Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, New York, USA. Department of Medicine, Weill Cornell Medical College, New York, New York, USA. Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, New York, USA. Ludwig Institute for Cancer Research, Lausanne, Switzerland. Department of Oncology, University of Lausanne, Lausanne, Switzerland. Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University, New York, New York, USA. 7 These authors contributed equally to this work. 8 Correspondence should be addressed to J.A.J. ( Received 9 March 7; accepted June 7; published online July 7; DOI:.8/ncb78 97 NATURE CELL BIOLOGY VOLUME 9 NUMBER 8 AUGUST 7 7 Macmillan Publishers Limited, part of Springer Nature. All rights reserved.

3 A RT I C L E S CDb + Gr + cells as predominantly hyper-segmented neutrophils (Supplementary Fig. b). Of note, we observed no changes in the proportions of bulk T lymphocytes (total CD + cells; 8 9% of total cells) or T-cell subsets (CD + CD +, % of total cells; CD + CD8 +, % of total cells) in lungs (Supplementary Fig. c). To uncouple whether this increase in lung neutrophils was due to high adiposity or diet content, we first analysed the genetic model of obesity, in which animals fed a normal diet exhibit rapid weight gain (Fig. c) due to hyperphagia secondary to leptin deficiency. Ob/ob lungs exhibited elevated proportions of neutrophils by flow cytometry, but no significant changes in overall leukocytes or macrophages (Fig. d). In a reciprocal experiment, we employed a BALB/c model of obesity resistance, whereby BALB/c mice were fed or diet for weeks, but did not gain weight (Fig. e). Unlike most other mouse strains, this obesity-resistance phenotype is inherent to BALB/c animals. We found no significant increase in neutrophils (Fig. f), in contrast to results from DIO and mice. These data suggest that the increase in lung neutrophils is due to high adiposity of obese animals, rather than diet/nutrient content. We next profiled other common organs for breast cancer dissemination, including liver and brain. DIO mice exhibited no change in immune cell proportions in brain, including macrophages and neutrophils (Supplementary Fig. d and Supplementary Table ). While we detected a significant increase in neutrophil proportions in the liver, these differences were inversely correlated with the model (Supplementary Fig. e), suggesting that the changes in this organ may be diet-dependent, unlike our findings in the lung (Fig. d). We therefore focused subsequent experiments on the effects of adiposity on lung inflammation, and neutrophils specifically, as this was the only immune cell population consistently elevated in the lung across the different obesity models. We next asked whether obesity-associated lung neutrophilia was reversible. Animals were enrolled on diet for weeks, and switched to for an additional 7 weeks (Supplementary Fig. f). Mice lost an average of 8% of their body weight over the diet-switch period (Fig. g). Lung neutrophils decreased after diet-switching, compared with animals that remained on continuous diet (Fig. h), indicating that obesity-associated lung neutrophilia is reversible with weight loss. Obesity-associated lung neutrophilia is accompanied by pro-metastatic gene expression changes and enhanced metastatic progression It is known that neutrophils, defined as either CDb + Gr +/hi or CDb + LyG + by flow cytometry, are elevated in the pre-metastatic lung in PyMT breast cancer models, and become activated to support metastatic progression,,7,9. We therefore asked whether lung neutrophils were activated in obese mice, and how this might affect breast cancer metastasis. We purified lung neutrophils from and animals (DIO model) using fluorescence-activated cell sorting (FACS), and performed quantitative PCR with reverse transcription (qrt PCR) for a panel of markers relevant to neutrophil biology in cancer. We focused on genes related to mobilization (Cxcl, Cxcl, Cxcr, Cxcr and Tlr ), N-like polarization (Arg, Ccl, Ccl and Tgfb,7 ), immune suppression (Il, Nox, Nos, Pge, Ptgs and Pdl 8 ) and activation/expansion (Ilb and Nlrp, Alox, SA9 and SA8 ). Markers of N-like polarization were significantly downregulated in neutrophils (Fig. a), suggesting discordance with the N/N-like paradigm. We observed significant increases in several markers associated with mobilization (/ upregulated) and activation (/ upregulated), and variable changes in markers associated with immune suppression (/ upregulated and / downregulated; Fig. a), indicating that neutrophils were phenotypically different from neutrophils, in addition to being more abundant. Interestingly, many gene expression changes in association with obesity were indicative of a pro-metastatic phenotype, for example, Alox, Sa8/9 and Cxcr/,,,. We next investigated whether the elevation in neutrophil number/activation during obesity was associated with enhanced lung metastasis. We generated a syngeneic immune-competent breast cancer model compatible with the BL DIO model, by isolating a panel of cell lines from BL MMTV-PyMT mice and confirming that they form orthotopic tumours that spontaneously metastasize to lung after months (Supplementary Fig. a d). Orthotopic transplantation of two of these lines (99LN, 8R) into the DIO model revealed that primary tumour growth was modestly enhanced with feeding over a -month period (Fig. b and Supplementary Fig. e). Systemic neutrophilia was monitored at different time points: prior to tumour cell injection ( d), when tumours were palpable ( d), during early phases of primary tumour growth (8 d; pre-metastatic niche) and late phases of primary tumour growth ( d; micro-metastatic disease). At d, circulating neutrophils were elevated in versus mice, and as primary tumours grew, a significant difference in neutrophilia was maintained (Fig. c). At early phases of primary tumour growth (8 d), we quantified neutrophils in the pre-metastatic lung, and found that while lean tumour-bearing mice exhibited lung neutrophilia compared with lean non-tumour-bearing mice (as has been reported,,7,9,7,, ), obesity significantly enhanced neutrophilia in the tumour-bearing setting (Fig. d). At the trial endpoint ( d), we found elevated micro-metastases in the lung (Fig. e and Supplementary Fig. f), concomitant with increased lung neutrophilia in obese compared with lean mice (Fig. d). These data indicate that obesity enhances spontaneous lung metastasis, and contributes to lung neutrophilia in a manner that is additive to pre-metastatic niche observations obtained in standard (lean) mouse models. Obesity enhances experimental breast cancer metastasis to lung in a neutrophil-dependent manner Given the differences in primary tumour volumes between and mice, we investigated whether obesity affects experimental metastasis of breast cancer cells via tail vein injection. We labelled 99LN cells with luciferase-gfp, to allow for bioluminescent imaging (BLI) and flow cytometry, and monitored experimental lung metastasis over weeks. mice exhibited significantly higher luminescence in the lung compared with mice after weeks (Fig. a,b), which was confirmed by flow cytometry for GFP + tumour cells (Fig. c). Moreover, we found that the increased proportion of neutrophils during obesity was also evident in the tumour-bearing setting (Fig. d). Given the roles of neutrophils in establishing a pre-metastatic niche,7,9 and influencing early seeding,, we examined the effects of obesity on earlier metastatic time points. We analysed lung metastasis 8 h following tail vein injection of 99LN cells into the DIO model. mice exhibited significantly higher luminescence in the 97 NATURE CELL BIOLOGY VOLUME 9 NUMBER 8 AUGUST 7 7 Macmillan Publishers Limited, part of Springer Nature. All rights reserved.

7 A RT I C L E S was capable of enhancing differentiation towards a CDb + Gr + phenotype (Fig. e). Antibody-mediated neutralization of GM-CSF in the context of serum reduced CDb + Gr + differentiation in vitro, to a level comparable to serum (Fig. f), demonstrating the necessity of this factor. Intriguingly, analysis of GM-CSF (Csf) expression by qrt PCR of bulk tissues from obese mice revealed a marked enrichment in whole lung compared with spleen, blood, BM, liver and fat (Fig. g and Supplementary Fig. e k). This is consistent with Csf mouse knockout studies demonstrating a tissue-specific role for GM-CSF in lung physiology and inflammation 7 9. Moreover, in FACS-purified cell populations from obese lungs, Csf was expressed by multiple immune cell types including various myeloid cells (for example, LyC hi monocytes), and CD + bulk T cells (Fig. h), suggesting that maintenance of neutrophils in the lung is due to both autocrine signalling and paracrine interactions with additional immune cell populations. These data implicate GM-CSF as a regulator of lung neutrophilia in the context of obesity. To test whether GM-CSF alone was capable of recapitulating obesity-induced lung neutrophilia in vivo, we treated animals with recombinant (r) GM-CSF for d, followed by flow cytometry analysis. Even after this short time period, there was a significant increase in the proportion of neutrophils in blood and lungs in response to rgm-csf compared with controls (Fig. a). This result is consistent with previous reports showing that GM-CSF regulates lung neutrophilia in models of pulmonary alveolar proteinosis, asthma, bronchitis and pulmonary infection 8,8. Significant differences in neutrophil numbers within immune reservoirs such as the spleen or BM were not observed during this short experimental time course (Supplementary Fig. a,b), suggesting that this change was not driven by neutrophil expansion within those depots. We next asked whether GM-CSF was sufficient to modulate metastatic outcome in vivo. We treated animals on a normal diet with rgm-csf for d to induce neutrophilia (Fig. a), followed by a 8 h experimental metastasis assay using 99LN breast cancer cells injected intravenously (Fig. b). We observed a significant increase in metastases, as measured by BLI in the lung at 8 h, in animals treated with rgm-csf compared with PBS controls (Fig. c). In a reciprocal experiment, we enrolled animals on either a or diet for weeks, followed by antibody-mediated neutralization of GM-CSF prior to injecting 99LN cells intravenously (Fig. d). At the trial endpoint of 8 h we found that GM-CSF neutralization in -fed animals, but not animals, resulted in a significant decrease in lung BLI compared with IgG control (Fig. e), corroborating our Gr-neutralization results (Fig. h,i). These results were validated by flow cytometry of GFP + tumour cells in lung (Fig. f). Importantly, when we immunoprofiled the lungs, we found that the proportion of neutrophils was significantly reduced in animals after GM-CSF neutralization compared with IgG, but this effect was not observed for animals (Fig. g). Together, these results demonstrate that obesity-associated GM-CSF is critical to support lung neutrophilia and enhanced metastatic seeding. We then asked whether serum GM-CSF was elevated in response to a primary tumour, as reported for the related factor G-CSF,7,9. Enzyme-linked immunosorbent assay (ELISA) for GM-CSF in serum isolated from the orthotopic experiments showed a modest (non-significant) increase in GM-CSF in lean mice in response to the presence of a primary tumour; this effect was significantly increased with obesity, and even more pronounced during later stages of progression (Fig. h). These data indicate that obesity may exacerbate the stimulatory effects of a primary tumour on neutrophils. IL signalling supports obesity-associated lung neutrophilia We next explored how obesity induces GM-CSF expression. From our findings in and Balb/c mice (Fig. c f), we determined that adiposity was an important contributor to lung neutrophilia. Therefore, we investigated which of the 8 upregulated factors from the cytokine array (Fig. d) were specifically enriched in adipose tissue. We found that Il expression was enriched in both subcutaneous and visceral fat in obese animals (Fig. a), and highest in adipose tissue compared with spleen, blood, BM, liver and lung (Supplementary Fig. e). IL plays a canonical role in regulating eosinophils, another type of granulocyte, during allergic asthma 9,. Given the link between obesity and adult-onset asthma 9, we wanted to determine the relevance of IL to neutrophil regulation. In animals, treatment with ril for d significantly increased the proportion of lung neutrophils compared with PBS controls (Fig. b,c). CD + T cells, another GM-CSF-producing cell population in the lung (Fig. h), were also elevated in response to ril (Supplementary Fig. 7a). Furthermore, increased neutrophils led to enhanced experimental lung metastasis, which was blocked by GM-CSF neutralization (Supplementary Fig. 7b d). These data suggest that neutrophils are in part regulated by IL, either by direct or indirect mechanisms. We next determined which cell types respond directly to IL, by identifying populations that express the IL receptor. The IL receptor has an alpha-subunit (ILrα) and beta-subunit (Csfrβ). Interestingly, the beta-subunit is shared between IL, GM-CSF and IL; a reflection of their complementary roles during airway inflammation and Th immunity in asthma. Csfrβ is broadly expressed on myeloid cells, and induces signal transduction in response to IL, while ILrα expression is more limited and dictates cellular specificity of IL signalling. Most of the literature on ILrαup is focused on expression by eosinophils (reviewed in ref. ), with only a few studies reporting eosinophil-independent effects of IL. However, by flow cytometry we determined that 99% of neutrophils and 8% of monocytes isolated from peripheral blood express ILrα, as compared with CDb + SSC hi SiglecF + eosinophils, which were % positive for ILrα as expected (Fig. d). When we quantified Csfra, Csfrb and Ilra expression in FACS-purified ILrα + lung neutrophils, eosinophils and monocytes from the DIO model, we found that in several instances, the genes encoding these receptor subunits were upregulated by obesity (Fig. e). This is consistent with reports of upregulated Ilra in neutrophils and monocytes in response to sepsis-associated inflammation, and methylation analyses that predicted ILRA upregulation in circulating immune cells in response to obesity in humans 7. Corroborating these results, by flow cytometry we found a subset of ILRα + intermediate monocytes from human blood (mean =.% positive), as well as a smaller proportion of neutrophils (mean =.% positive), classical monocytes (mean =.% positive), and non-classical monocytes (mean =.8% positive) when eosinophils were used as a positive gating control (Fig. f) Macmillan Publishers Limited, part of Springer Nature. All rights reserved. NATURE CELL BIOLOGY VOLUME 9 NUMBER 8 AUGUST 7

10 A RT I C L E S We were unable to detect Ilra expression by qrt PCR on CD + T cells isolated from lungs in the DIO model. However, given our findings that CD + T cells isolated from obese lung tissue express Csf (Fig. h), and the increase in CD + T-cell numbers in the lung following ril treatment in vivo (Supplementary Fig. 7a), we wanted to directly test whether T cells were important for IL- mediated lung neutrophilia. We treated athymic nude mice with ril for d, and found that lung neutrophilia in response to IL treatment did not occur in the absence of T cells (Supplementary Fig. 7e). In addition, the proportion of ILrα + myeloid cells in nude mice was significantly reduced compared with BL mice (monocytes reduced from 8% to <%; neutrophils 99% to 9%; eosinophils % to 9%; Supplementary Fig. 7f). Similar results were observed in NOD-scid Ilrg null (NSG) mice, which lack mature T, B and NK cells (Supplementary Fig. 7e,f). Taken together, our findings indicate that T cells are indispensable for IL-induced lung neutrophilia. We next asked which ILrα + cell type was capable of mediating a functional response to IL. We FACS-purified ILrα + cells from the peripheral blood of BL mice, including monocytes, neutrophils and eosinophils, and treated them in vitro with ril. Not surprisingly, by flow cytometry we observed increased Ki7 + proliferating ILrα + eosinophils in response to ril (Fig. g) a canonical effect of IL in this cell type. We also found increased proliferation of ILrα + monocytes in response to ril in vitro (Fig. g), concomitant with upregulated Csf expression by qrt PCR (Fig. h). In the DIO model, we confirmed that obesity increases ILrα + monocyte abundance in blood and lung, concomitant with an increase in Csf expression, compared with controls (Fig. i,j). These data indicate that ILrα + monocytes directly respond to IL by expanding in numbers and producing GM-CSF, and may thereby contribute to lung neutrophilia during obesity. Obesity enhances lung homing of neutrophils in an IL-dependent manner Given that lung neutrophils upregulate genes associated with mobilization (Fig. a), such as Tlr, Cxcr and Cxcr, we asked whether obesity alters homing and/or retention of neutrophils in the lung, and whether this is dependent on IL. We isolated and fluorescently labelled neutrophils from BM of (green) and (red) donors, mixed them :, and performed adoptive cell transfer back into, or + anti-il recipients (Fig. 7a,b). We found that h post-adoptive transfer, the majority of labelled neutrophils in the lung across all recipient groups were from donors (Fig. 7c,d), despite equivalent proportions of and neutrophils in blood (Fig. 7e), indicating that neutrophils from obese animals are predisposed to lung homing. Furthermore, lung trafficking of donor neutrophils was significantly enhanced in recipients compared with recipients, and this effect was mitigated by IL neutralization (Fig. 7c). This suggests that obesity primes the lung microenvironment to support neutrophil homing in an IL-dependent manner. After 8 h, neutrophil clearance within the lungs and blood was comparable between donor groups (Supplementary Fig. 8a,b); however, recipient animals retained the highest levels of lung neutrophils regardless of donor genotype (Fig. 7f,g). Interestingly, this capacity for neutrophil retention was blocked by IL neutralization (Fig. 7f,g). These data indicate that obesity supports increased lung homing and retention of neutrophils in an IL-dependent manner. Weight loss reduces obesity-associated lung neutrophilia and metastasis Given our observation that diet-switch reverses obesity-induced lung neutrophilia in the DIO model (Fig. g,h), and the relevance of this non-invasive intervention strategy to patients, we investigated whether weight loss could similarly reduce obesity-associated metastasis. We confirmed that ILrα + neutrophils and monocytes were upregulated in lungs, which was reversed by diet-switching (Fig. 8a c). We found that diet-switching reversed experimental lung metastasis (Fig. 8d), and the expression of key markers of neutrophil activation/mobilization, including Cxcr and SA8 (Fig. 8e). Finally, serum analysis by ELISA showed reduced GM-CSF and IL in diet-switched mice, compared with mice fed continuous diet (Fig. 8f). These data indicate that diet changes in association with weight loss may be sufficient to reverse the pro-metastatic effects of obesity. Finally, we asked whether weight loss intervention was sufficient to reduce key serum factors identified here (that is, GM-CSF, IL) in a human clinical trial. We obtained matched serum samples from female obese humans before and after % weight loss by caloric restriction 8 (ClinicalTrials.gov Identifier: NCT999). Consistent with results from the DIO model, there was a significant decrease in circulating neutrophils with weight loss 8, corroborating observations of reduced leukocytosis following bariatric surgery in obese humans 9. Following assessment of serum levels of GM-CSF and IL in matched individuals, we found that these factors were generally reduced following weight loss (Fig. 8g), Similarly, weight loss resulting from surgical intervention has been shown to reduce serum levels of GM-CSF and IL in accordance with a reduction in BMI. These results demonstrate the potential translational relevance of our findings to humans, and indicate that weight loss interventions may be useful to improve outcomes in obese breast cancer patients. DISCUSSION In this study, we have identified lung neutrophilia as a secondary effect of obesity. This occurs independently of diet; rather it is directly related to increased adiposity and the expression of IL by adipose tissue. IL has been reported as a negative regulator of adipose tissue expansion and insulin resistance, thereby working to constrain the effects of obesity on metabolism. While in adipose tissue IL supports homeostasis, the increase in serum IL has the opposite effect on lung. We found that IL increases Csf expression by ILrα + monocytes, and neutrophil trafficking to lung. Furthermore, increased serum GM-CSF promotes myelopoiesis, leading to an expansion of peripheral neutrophils (Fig. 8h). Collectively these findings demonstrate that obesity causes lung inflammation, despite the lack of adipose tissue in this organ, by acting through the systemic environment. In tumour models, we showed that obesity-associated lung neutrophilia enhances breast cancer metastasis to this organ, and that depletion of Gr + cells in obese animals reverses this effect. This is reminiscent of the pre-metastatic niche concept, although in this case the elevation of pro-metastatic neutrophils is observed NATURE CELL BIOLOGY VOLUME 9 NUMBER 8 AUGUST 7 7 Macmillan Publishers Limited, part of Springer Nature. All rights reserved. 98

3 RESULTS As outlined under External contributions (see appendix 7.1), the group of Prof. Gröne at the DKFZ in Heidelberg (Dept. of Cellular and Molecular pathology) contributed to this work by performing

Supplementary Figure 1. Example of gating strategy Legend Supplementary Figure 1: First, gating is performed to include only single cells (singlets) (A) and CD3+ cells (B). After gating on the lymphocyte

This is an by copyright after embargo allowed publisher s PDF of an article published in Jablonska, J., Leschner, S., Westphal, K., Lienenklaus, S., Weiss, S. Neutrophils responsive to endogenous IFN-β

Interactions between innate immunity & adaptive immunity What happens to T cells after they leave the thymus? Naïve T cells exit the thymus and enter the bloodstream. If they remain in the bloodstream,

Examples of questions for Cellular Immunology/Cellular Biology and Immunology Each student gets a set of 6 questions, so that each set contains different types of questions and that the set of questions

LECTURE: 07 Title: IMMUNE CELL SURFACE RECEPTORS AND THEIR FUNCTIONS LEARNING OBJECTIVES: The student should be able to: The chemical nature of the cellular surface receptors. Define the location of the

Effect of Early onset Obesity versus Late onset Obesity on Immune Cells in Regional Adipose Tissue Depots: A Pilot Study Vi Dam A Thesis in The Department of Exercise Science Presented in Partial Fulfillment

Bead Based Assays for Cytokine Detection September 27, 2014 6 th EFIS-EJI South East European Immunology School SEEIS 2014 Timisoara, Romania The Cells of the Immune System The Immune Reaction (Th2) (Th1)

Supplementary Fig. S1. Evaluation of the purity and maturation of macrophage cultures tested by flow cytometry. The lymphocytic/monocytic cellular fraction was isolated from buffy coats of healthy donors

INCUCYTE LIVE-CELL ANALYSIS SYSTEM Assays for Immuno-oncology Research Real-time automated measurements of immune and tumor cell dynamics within your incubator See what your cells are doing and when they

Autoimmunity Autoimmunity arises because of defects in central or peripheral tolerance of lymphocytes to selfantigens Autoimmune disease can be caused to primary defects in B cells, T cells and possibly

Posters and Presentations June 2017: American Society of Clinical Oncology (ASCO) Annual - Preliminary Correlative Analysis of PD-L1 expression from the SUNRISE Study. View April 2017: American Association

Washington University School of Medicine Digital Commons@Becker Open Access Publications 2015 mtor and MEK1/2 inhibition differentially modulate tumor growth and the immune microenvironment in syngeneic

1 The immune system The immune response The immune system comprises two arms functioning cooperatively to provide a comprehensive protective response: the innate and the adaptive immune system. The innate

1 International Graduate Research Programme in Cardiovascular Science This work has been supported by the European Community s Sixth Framework Programme under grant agreement n LSHM-CT-2005-01883 EUGeneHeart.

Role of microenvironment and tumor interactions in melanoma progression with special regard to the prognostic significance of immune cell infiltrate PhD thesis Dr. Anita Mohos Doctoral School of Pathological

The immune response against cancer Maries van den Broek Institute of Experimental Immunology vandenbroek@immunology.uzh.ch The immune system Main cells of the immune system Dendritic cell Monocyte Macrophage

The Hallmarks of Cancer This illustration encompasses the six hallmark capabilities originally proposed in our 2000 perspective. The past decade has witnessed remarkable progress toward understanding the